Hepatic transcriptome analysis identifies genes, polymorphisms and pathways involved in the fatty acids metabolism in sheep

Fatty acids (FA) in ruminants, especially unsaturated FA (USFA) have important impact in meat quality, nutritional value, and flavour quality of meat, and on consumer’s health. Identification of the genetic factors controlling the FA composition and metabolism is pivotal to select sheep that produce higher USFA and lower saturated (SFA) for the benefit of sheep industry and consumers. Therefore, this study was aimed to investigate the transcriptome profiling in the liver tissues collected from sheep with divergent USFA content in longissimus muscle using RNA deep-sequencing. From sheep (n = 100) population, liver tissues with higher (n = 3) and lower (n = 3) USFA content were analysed using Illumina HiSeq 2500. The total number of reads produced for each liver sample were ranged from 21.28 to 28.51 million with a median of 23.90 million. Approximately, 198 genes were differentially regulated with significance level of p-adjusted value <0.05. Among them, 100 genes were up-regulated, and 98 were down-regulated (p<0.01, FC>1.5) in the higher USFA group. A large proportion of key genes involved in FA biosynthesis, adipogenesis, fat deposition, and lipid metabolism were identified, such as APOA5, SLC25A30, GFPT1, LEPR, TGFBR2, FABP7, GSTCD, and CYP17A. Pathway analysis revealed that glycosaminoglycan biosynthesis- keratan sulfate, adipokine signaling, galactose metabolism, endocrine and other factors-regulating calcium metabolism, mineral metabolism, and PPAR signaling pathway were playing important regulatory roles in FA metabolism. Importantly, polymorphism and association analyses showed that mutation in APOA5, CFHR5, TGFBR2 and LEPR genes could be potential markers for the FA composition in sheep. These polymorphisms and transcriptome networks controlling the FA variation could be used as genetic markers for FA composition-related traits improvement. However, functional validation is required to confirm the effect of these SNPs in other sheep population in order to incorporate them in the sheep breeding program.

Prospective study for commercial and low-cost hyperspectral imaging systems to evaluate thermal tissue effect on bovine liver samples

Thermal ablation modalities, for example radiofrequency ablation (RFA) and microwave ablation, are intended to prompt controlled tumour removal by raising tissue temperature. However, monitoring the size of the resulting tissue damage during the thermal removal procedures is a challenging task. The objective of this study was to evaluate the observation of RFA on an ex vivo liver sample with both a commercial and a low-cost system to distinguish between the normal and the ablated regions as well as the thermally affected regions. RFA trials were conducted on five different ex vivo normal bovine samples and monitored initially by a custom hyperspectral (HS) camera to measure the diffuse reflectance (Rd) utilising a polychromatic light source (tungsten halogen lamp) within the spectral range 348–950 nm. Next, the light source was replaced with monochromatic LEDs (415, 565 and 660 nm) and a commercial charge-coupled device (CCD) camera was used instead of the HS camera. The system algorithm comprises image enhancement (normalisation and moving average filter) and image segmentation with K-means clustering, combining spectral and spatial information to assess the variable responses to polychromatic light and monochromatic LEDs to highlight the differences in the Rd properties of thermally affected/normal tissue regions. The measured spectral signatures of the various regions, besides the calculation of the standard deviations (δ) between the generated six groups, guided us to select three optimal wavelengths (420, 540 and 660 nm) to discriminate between these various regions. Next, we selected six spectral images to apply the image processing to (at 450, 500, 550, 600, 650 and 700 nm). We noticed that the optimum image is the superimposed spectral images at 550, 600, 650 and 700 nm, which are capable of discriminating between the various regions. Later, we measured Rd with the CCD camera and commercially available monochromatic LED light sources at 415, 565 and 660 nm. Compared to the HS camera results, this system was more capable of identifying the ablated and the thermally affected regions of surface RFA than the side-penetration RFA of the investigated ex vivo liver samples. However, we succeeded in developing a low-cost system that provides satisfactory information to highlight the ablated and thermally affected region to improve the outcome of surgical tumour ablation with much shorter time for image capture and processing compared to the HS system.

Case Report: Gastrointestinal Basidiobolomycosis with Multi-Organ Involvement Presented with Intussusception

Gastrointestinal basidiobolomycosis (GIB) is a rare, life-threatening fungal infection affecting immunocompetent individuals in tropical and subtropical regions. A diverse presentation of GIB has been reported, but no report has yet been published on intussusception. We describe a 23-month-old immunocompetent boy from a subtropical area in Iran who presented with intussusception. Prolonged fever, an abdominal mass, hepatomegaly, high erythrocyte sedimentation rate, and peripheral eosinophilia strongly suggested GIB. Accordingly, GIB was diagnosed based on the characteristic histopathology (the Splendore-Hoeppli phenomenon) detected in a liver sample taken via biopsy. Exploratory laparotomy showed several organs, including the colon, gall bladder, liver, and abdominal wall, were involved. Antifungal therapy with trimethoprim/sulfamethoxazole, liposomal amphotericin B, a saturated solution of potassium iodide, and surgical resection of involved tissues were used with improved outcome. The presence of non-septate fungal hyphal elements in the colonic mucosa led to the thickening of the bowel wall, leading to secondary intussusception.

Optical Signature Analysis of Liver Ablation Stages Exploiting Spatio-Spectral Imaging

Background and Objective: Thermal ablation modalities such as Radiofrequency ablation (RFA) / Microwave ablation (MWA) are deliberately used for marginally invasive tumor removal by escalating tissue temperature. For precise tumor extinguish, thermal ablation outcomes need routine monitoring for tissue necrosis in a challenging research task. The study aims to exploit hyperspectral imaging (HSI) to evaluate the impact of the liver tissue ablation. Materials and Methods: RFA with temperature range (≥80 °C) was accomplished on the ex vivo animal liver and evaluated using a spectral camera (400~1000 nm). The spectral signatures were extracted from the HSI data after the following processing steps: capturing three spectral data cubes for each liver sample with total 7-samples (before ablation, after ablation, and after ablation with sample slicing) using an HSI optical configuration. The custom HSI processing comprises “Top-hat and Bottom-hat transform” combined with “watershed transform” image segmentation to increase the intensity for a region of interest (ROI) of the investigated tissue, linking spectral and spatial data. Additionally, statistical analysis for HSI data was performed to exclusively select the best spectral band that discriminates between the normal, thermally-damaged, and ablated liver regions. Results: The variation of the optical parameters for the investigated liver samples provides variable interaction with the light diffuse reflection (Ŗd) over the spectrum range (400~1000 nm). Where, the extracting spectral information of the various tissue zones from the induced RFA linked to the hemoglobin, methemoglobin, and water permits variations. The generated spectral image after image enhancement utilizing “Top-hat and Bottom-hat transform” followed by “watershed segmentation”, showed high contrast between normal and thermal regions at a wavelength (600 nm). However, the wavelength (900 nm) shows a high variance between the normal and ablated regions. Finally, delineation of the thermal and ablated regions on the complemented enhanced image. Conclusion: HSI is considered a promising optical noninvasive technique for monitoring the RFA toward enhancing the ablation-based treatment for liver tumor outcomes.

Hematology Profile and Liver Histopathology in Escherichia coli Infected Layers Treated with Combination of Phyllanthus ( Phyllanthus niruri L. ) and Turmeric ( Curcuma domestica )

Colibacillosis a disease that can cause considerable economic loss, remains an important health problem. Phyllanthus (Phyllanthus niruri L) and turmeric (Curcuma domestica) are herbs that can be used as immunomodulators. This study was aimed to determine the level of safety of the combination of phyllanthus and turmeric on hematology profile and liver histopathology of layers with colibacillosis. The layers were assigned to the following of 5 groups: a) colibacillosis group without treatment, b) colibacillosis group with 500 mg/kg BW of phyllanthus, c) colibacillosis group with 300 mg/kg BW of turmeric, d) colibacillosis group with phyllanthus and turmeric combination (1:1), e) colibacillosis group with combination of phyllanthus and turmeric (1:2) . After 21 days of treatment, blood and liver sample were collected. The hematological profile (hemoglobin, hematocrit, erythrocyte and leukocyte counts) and liver histology were examined. The result were analyzed statistically by one-way ANOVA. The group that received phyllanthus had higher levels of hemoglobine, haematocrit and erythrocytes than the control group. However, no significant differences were found for the overall groups. Treatment with the combination of turmeric and phyllanthus for 21 days did not cause changes in the hematological profiles or liver histology, and therefor this herbal combination can be used as an alternative therapy for colibacillosis in layers.

Emergence of fowl aviadenovirus C-4 in a backyard chicken flock in California

Fowl aviadenovirus (FAdV) species D and E are associated with inclusion body hepatitis (IBH); species C, serotype 4 (hereafter, FAdV4) is associated with hepatitis–hydropericardium syndrome (HHS) in young chickens. Outbreaks of HHS have led to significant losses in the poultry industry in several countries, predominantly in China. In April 2020, FAdV4 was detected in a remote backyard flock in California. In a mixed flock of chickens of various breeds and ages (6 mo to 2 y old), 7 of 30 were found dead within a week without premonitory signs. One additional bird died after the flock was relocated to fresh pasture, bringing the total mortality to 8 of 30 (27%). Postmortem examination of 3 birds revealed good body condition scores and active laying. One chicken had subtle hemorrhages throughout the liver, and the other 2 had diffusely dark mahogany livers. On histopathology, 2 chickens had hepatic necrosis with hepatocytes containing large, mostly basophilic, intranuclear inclusion bodies, identified by electron microscopy as 82.2-nm diameter adenoviral particles. Virus isolation and genomic sequencing performed on a liver sample revealed strains with 99.9% homology to FAdV4 isolates reported from China. To our knowledge, FAdV4 has not been reported in the United States to date. Furthermore, the chickens affected here were all adults and exhibited a variation of serotype 4 disease in which IBH was present but not hydropericardium.

The Occurrence of Heavy Metals (Cadmium and Lead) in the Liver of Hogs in the Region of Vojvodina

Summary The purpose of this research is to investigate the occurrence of two heavy metals (namely cadmium (Cd) and lead (Pb)) in the liver of hogs bred in different locations in Vojvodina. A total of 30 liver samples were collected from ten pig farms in Vojvodina for experimental purposes in the period from December 2017 to January 2018. The samples collected were analysed for the presence of lead and cadmium. The average concentration of lead in all the samples was 0.39 mg/kg wet weight, whereas the samples from only one farm of the ten considered were found to contain a slightly higher average lead concentration than permitted by the Serbian standard. A lead concentration of 0.86 mg/kg wet weight was detected in one liver sample from this farm. The maximum permitted lead concentration was exceeded in the liver samples obtained from three farms. However, all the liver samples analysed were found to contain the permitted levels of cadmium, with an average cadmium concentration of 0.12 mg/kg wet weight and a maximum cadmium concentration of 0.48 mg/kg wet weight. The occurrence of heavy metals and their origin in the pig’s offals should be examined in greater detail in future research, especially because pig’s offals are used in the meat processing industry.

Thin layer chromatographic detection of colistin sulfate antibiotic residues in poultry tissues

Residues of antibiotic drugs in food and food products have been received much attention in recent years because of growing food safety concerns for public health. There are serious effects of antibiotic residues in meat for human consumption (e.g., increasing antimicrobial resistance, carcinogenicity, mutagenicity, and hypersensitivity). In this study, we used broiler chicks as a laboratory animal to detect the colistin sulfate antibiotic residue for human health concerns. The day-old broiler chicks were collected and reared for up to 31 days. The treatment was started from the day 16th until sacrifice. The chicks were randomly divided into three groups namely control group (Group A), discriminate antibiotic group (Group B) and indiscriminate antibiotic group (Group C) on the 14th day. The discriminate group was treated with an antibiotic, colistin sulfate maintaining the withdrawal period of one week. In case of indiscriminate group the withdrawal period was not maintained and antibiotic treatment was continued till the day of sacrifice. The body weights of the birds were recorded daily. The mean body weight was highest in indiscriminate group (1261.15±16.37gm) followed by discriminate group (1156.15±18.23gm) and the lowest was in control group (1008.49±18.11gm). The differences among mean weight gain were statistically significant (P<0.05) in antibiotic treated group compared with control group. The Thin Layer Chromatography revealed that all the samples were positive in indiscriminate group and in case of Liver, Kidney and Spleen, it was 100%. In case of discriminate antibiotic group all the samples were positive except thigh and breast muscle and the percentage was highest in Liver sample (66.67%). There was no positive sample in control group. The results were statistically significant (P<0.05). From the above findings, this research could be considered a need based research in Bangladesh to ascertain the influential effect of antibiotic abuse in poultry industry.

Heroin Fatality in a Feline: A Case Report with Postmortem Liver Concentrations

A case of feline intoxication and fatality with the illicit drug heroin is described. A 5-year-old castrated male domestic shorthair cat was recently diagnosed with an active pneumonitis and left at home for a couple of days under the care of another resident. Upon return, the owner found his cat dead with strong suspicion of foul play. The cat was necropsied by a local veterinary clinic to retrieve the liver for diagnostic toxicology. The postmortem liver sample screened positive for 6-acetylmorphine and 6-acetylcodeine by gas chromatography mass spectrometry. Deconvolution techniques were applied to chromatograms, which revealed the additional presence of morphine and mirtazapine. Subsequent quantitation of mirtazapine, heroin, morphine, 6-acetylmorphine and 6-acetylcodeine was performed by gas chromatography tandem quadrupole mass spectrometry. Although companion animal fatalities arising from toxicities are a likely consequence of drug abuse in a home, this is the first reported case of a malicious feline fatality resulting from heroin with quantitation of heroin metabolites.

Postmortem Changes in pH, Color, Drip Loss, and Non-Protein Nitrogen in Beef Liver and Lungs During Storage in Refrigerator

Beef offal are consumed by people in some countries specifically in Asia. Beef liver and lungs are favorite food which are used as meat in traditional food. The objective of this study was to determine the postmortem changes in pH, color, drip loss, and non-protein nitrogen (NPN) content in beef liver and lungs during storage in refrigerator (3-4 ºC) until 5 d (120 h) after slaughter. The beef liver and lungs were collected from the abattoir and transported in cool box (<7 ºC) to the laboratory within 3 hours. The samples size of beef liver and lungs were 20 for each observation time. In the laboratory the beef liver and lungs were measured directly for pH value, color (L*, a*, and b*), drip loss, and NPN content at 4 h postmortem (pm) and afterwards every beef liver sample was sliced into 5 pieces of 100-120 g and stored in chiller of 3-4 ºC. The measurement of pH, color (L*, a*, and b* values), drip loss, and NPN content were conducted at 4 h, 24 h, 48 h, 72 h, 96 h, and 120 h postmortem. Data were analyzed descriptively and by comparing the 95% confidence interval of mean of each observation. The results showed that pH, color, drip loss, and NPN content in beef lungs were higher than the values in beef liver. The pH of beef liver and lungs declines until 96 h pm and 48 h pm, respectively. The L*, a*, and b* values of beef liver and lungs increased in general during storage. Drip loss and NPN in beef liver and lungs tended to increase significantly during storage. From this study the pH value and NPN can be used to determine the freshness of beef liver and lungs.